Three new endophytic Apiospora species (Apiosporaceae, Amphisphaeriales) from China.

Apiospora species are widely distributed fungi with diverse lifestyles, primarily functioning as plant pathogens, as well as exhibiting saprophytic and endophytic behaviors. This study reports the discovery of three new species of Apiospora, namely A.gongcheniae, A.paragongcheniae, and A.neogongcheniae, isolated from healthy Poaceae plants in China. These novel species were identified through a multi-gene phylogenetic analysis. The phylogenetic analysis of the combined ITS, LSU, tef1, and tub2 sequence data revealed that the three new species formed a robustly supported clade with A.garethjonesii, A.neogarethjonesii, A.setostroma, A.subrosea, A.mytilomorpha, and A.neobambusae. Detailed descriptions of the newly discovered species are provided and compared with closely related species to enhance our understanding of the genus Apiospora.

First Report of Neofusicoccum parvum Causing Seedpod Blight on Lotus (Nelumbo nucifera) in China.

Lotus (Nelumbo nucifera Gaertn.) is a widely cultivated plant in China, and the fruit lotus variety has a high economic value attributed to the exquisite flavor of its fresh seeds. During the summer of 2023, an unidentified blight was observed affecting lotus seedpods in Jiande City, Zhejiang province, with approximately 65% of seedpods impacted in a 130-hectare area. The initial symptoms included dark purple spots on the lotus seedpod surface, which gradually expanded over time. After 5 to 7 days, the entire seedpod turned black, withering, and rendering the lotus seeds inedible. To identify the causal agent, tissues from symptomatic seedpods were excised and disinfected in 75% ethanol for 60 s, and washed twice in sterile distilled water. The disinfected symptomatic tissues (5 × 5 mm) were plated on potato dextrose agar (PDA), incubated at 25 ℃, transferred hyphal tips to obtain pure isolates after 3 days. Fungal colonies exhibiting Botryosphaeriaceae morphology were isolated from 33% of the samples (n = 15). Pure cultures were grown on PDA for both morphological and molecular identification. The colonies displayed a white aerial mycelium, turning olivaceous grey after 7 days. Pycnidia were produced within 3 weeks on PDA with added sterilized healthy lotus seedpod pieces on the surface. Conidia were hyaline, unicellular, ellipsoidal, 12.65 to 20.72 × 3.92 to 9.38 µm in size (mean 16.67 × 6.24 µm, n = 100). To determine the fungal species, genomic DNA was extracted from one representative isolate (ZJUP1112-1), to amplify four gene loci through polymerase chain reactions (PCR): rDNA internal transcribed spacer (ITS) with primers ITS1/ITS4, rDNA large subunit (LSU) with LR0R/LR5, the translation elongation factor 1-alpha gene (tef1) with EF1-728F/EF1-986R, and ß-tubulin gene (tub2) with Bt2a/Bt2b. The PCR products were Sanger sequenced in Zhejiang Shangya biotechnology co., LTD, and the resulting sequences were assembled and deposited in GenBank (ITS: OR740546; LSU: OR740547; tef1: OR776996; tub2: OR776997). BLAST searches indicated the highest nucleotide sequence identity with the reference strains of Neofusicoccum parvum CMW 9081 (ITS: 98.8%, AY236943; LSU: 100%, AY928045; tef1: 99.6%, AY236888; tub2: 99.3%, AY236917). Multi-locus phylogenetic analyses revealed that isolate ZJUP1112-1 formed a highly supported clade with N. parvum. Pathogenicity tests were performed on healthy lotus seedpods using mycelial plugs (5 mm diameter) from actively growing colonies of ZJUP1112-1 that were placed onto the front and side of the seedpods (6 each). Controls received PDA plugs. Treated seedpods were wrapped with parafilm and incubated at 25 ℃ and the experiment was repeated three times. After 5 days, dark purple lesions were observed on the inoculated seedpods, whereas controls remained symptomless. The same isolate was recovered from the margin of resulting lesions and confirmed by morphology, thus fulfilling Koch's postulates. N. parvum is a polyphagous pathogen causing blights and fruit rot on multiple economically important fruit crops, such as cacao (Puig et al. 2019), walnut (Chen et al. 2019), pistachio (Lopez-Moral et al. 2020), chestnut (Seddaiu et al. 2021), blueberry (Spetik et al. 2023) and mango (Polizzi et al. 2022), among others. To the best of our knowledge, this is the first report of N. parvum causing seedpod blight on lotus seedpods in China, which contributes to a better understanding of the pathogens affecting this plant species in China.

First Report of Monosporascus cannonballus causing Root Rot and Vine Decline in Watermelon (Citrullus lanatus) in China.

In June 2023, a sudden outbreak root rot and vine decline symptoms was observed during a watermelon (Citrullus lanatus T.) variety demonstration trial located in Taizhou City, Zhejiang Province, China, with an incidence rate ranging from 75% to 100% and an affected area of nearly 2,000 square meters. The disease initially appeared with a rapid and alarming invasion of root rot and vine decline symptoms within watermelon plants. Affected plants exhibited rapid deterioration, showing symptoms of wilting, yellowing and eventual demise, predominantly during the pre-harvest stage. Notably, numerous black, spherical, erumpent perithecia were clearly visible on the watermelon's root epidermis, a characteristic trait of the disease. Symptomatic plant samples were rigorously disinfected with 75% ethanol, and plated on potato dextrose agar medium for incubation at 25°C, successfully isolate two potential strains. These isolates were inoculated in oatmeal agar and incubated in a 25℃ light incubator. After 30 days, mature perithecia, the same as those found on the watermelon's root epidermis, reached a diameter of 500 µm. Each perithecium contained several pear-shaped asci, 56 to 108.5 µm in length and 30.5 to 46.4 µm in width, typically holding 1, rarely 2 ascospores. These characteristics align precisely with the typical strains of Monosporascus cannonballus Pollack and Uecker (1974). Additionally, sequencing the internal transcribed spacer region of ribosomal DNA (ITS) gene (White et al., 1990), large subunit ribosomal RNA (LSU) gene (Rehner and Samuels 1995), and beta-tubulin (TUB) gene (Glass and Donaldson, 1995) were performed. BLAST analysis indicated the highest nucleotide sequence identity with M. cannonballus CBS 586.93 reference sequence (ITS: 100%, JQ771930; TUB: 98.99%, JQ907292). Representative sequences of isolate ZJUP0990-2 from these regions were deposited in GenBank (Accession No.: OR357656 for ITS; OR474500 for LSU; OR365762 for TUB). A multigene phylogenomic analysis (ITS-LSU-TUB) was undertaken to ascertain the exact phylogenetic position of M. cannonballus within the genus Monosporascus. The amalgamation of both morphological and molecular insights consistently reaffirmed the accurate classification of the causative agent as M. cannonballus. To validate the pathogenicity of M. cannonballus, a controlled greenhouse experiment was conducted using watermelon (cv. Nabite) as the subject. Mycelium fragments, harvested from the edge of the colony ZJUP0990-2, were inoculated into oat liquid medium and cultivated under dark conditions at a consistent temperature of 30°C for 7 days. After 20 days, the inoculated plants exhibited root rot and wilting, mirroring the symptoms observed during the field outbreak. In contrast, the control plants did not exhibit any signs of disease. M. cannonballus was successfully re-isolated from the symptomatic roots of the inoculated plants, satisfying Koch's postulates. This experiment was repeated three times. This pathogenic fungus has previously been documented as a menace to melons in various regions including Mexico (Chew-Madinaveitia et al., 2012) and Brazil (Sales et al., 2004), as well as watermelons in Brazil (Sales et al., 2010), northern Mexico (Gaytan-Mascorro et al., 2012), and Saudi Arabia (Karlatti et al., 1997). To our knowledge, this is the first reported presence of M. cannonballus on watermelons in China. This new disease poses a serious threat to watermelon production, potentially leading to severe economic losses and impacting food security.

First report of Trunk Canker Caused by Botryosphaeria dothidea on Cinnamomum camphora in China.

Cinnamomum camphora, an essential woody plant in China, experienced a severe outbreak of trunk canker affecting an area of 540,000 square meters in the Xiaoshan District of Zhejiang province during the early summer of 2022. The observed symptoms included stem canker, dieback, twig blight, and extensive vascular discoloration, with an incidence rate ranging from 45% to 70%. To investigate the etiology of the disease, symptomatic plant samples were subjected to rigorous surface sterilization involving washing with running tap water, followed by surface sterilization using 75% ethanol. The samples were then rinsed twice with sterile distilled water, plated on potato dextrose agar (PDA) medium, and incubated at a temperature of 28°C. The isolated fungi exhibited characteristics consistent with the species Botryosphaeria dothidea. Fungal isolates displayed abundant white aerial mycelium, which darkened to grey after five to seven days, eventually giving rise to black pycnidia. Single hyphal tip cultures of putative two isolates were stored at the Agricultural Experiment Station of Zhejiang University. Conidia formed on pycnidia were one-celled, hyaline, aseptate, and fusiform, with dimensions of 18.34-27.12 µm x 3.77-6.84 µm (average 22.90 µm x 5.20 µm) (one hundred conidia were measured). To determine the fungal species, genomic DNA was extracted from individual isolates ZJUP0868, and subjected to DNA sequence analysis of four gene regions: the internal transcribed spacer (ITS) using the primer-pair ITS4 and ITS1 (White et al., 1990), the large subunit (LSU) gene with LR0R and LR5 (Rehner and Samuels, 1995), the small subunit (SSU) gene with SR1R and SR7 (Zoller et al., 1999), and the translation elongation factor 1-alpha (tef1) with EF1-983F and EF1-2218R (Rehner and Buckley, 2005). Furthermore, DNA sequence analysis of above four genes was performed. BLAST analysis indicated the highest nucleotide sequence identity with Botryosphaeria dothidea CBS 115476 reference sequence (ITS: 99.59%, KF766151; LSU: 99.88%, DQ377852; SSU: 100%, NG_062738; tef1: 98.93%, AY236898). Representative sequences of isolate ZJUP0868 from these regions were deposited in GenBank (ITS: Accession No. OR192838; LSU: Accession No:OR248147; SSU: Accession No:OR248174; tef1: Accession No. OR262053). A phylogenomic analysis was conducted to determine the phylogenetic position of Botryosphaeria dothidea in the family Botryosphaeriaceae. The combined morphological and molecular findings confirmed the identification of the pathogen as Botryosphaeria dothidea. Pathogenicity tests were conducted by stem inoculation of two-year-old Cinnamomum camphora baranches. Mycelial plugs (2-3 mm in diameter) from actively growing colonies of B. dothidea (PDA) were applied to bark of similar size on the middle point of the stems. Inoculated barks were wrapped with Parafilm, while control branches received sterile PDA plugs. Inoculated and control branches (7 each) were maintained in a greenhouse at 28°C. After two weeks, all inoculated plants exhibited dark vascular stem tissue, whereas the control plants remained healthy. B. dothidea was re-isolated from symptomatic tissues, thus fulfilling Koch's postulates. No symptoms were observed in the control branched, and B. dothidea was not re-isolated from their tissues. B. dothidea , the type species of Botryosphaeria (Botryosphaeriaceae, Botryosphaeriales), is commonly associated with cankers and dieback in woody plants. Previous reports have identified Botryosphaeria dothidea as a pathogen causing stem dieback and trunk canker on Glycine max (Chen et al., 2020), Camellia oleifera (Hao et al., 2022), and Gleditsia sinensis (Huang et al., 2020). Additionally, B. dothidea has been reported to cause leave wilt on various plant species in China, including Daimyo oak (Liu et al., 2023) and Cornus officinalis (Zhang et al., 2022). To the best of our knowledge, this is the first report of B. dothidea on C. camphora in China. This findings contribute to a better understanding of the pathogens affecting Cinnamomum camphora in the China.

Clinical impact of blood pressure on cardiovascular death in patients 80 years and older following acute myocardial infarction: a prospective cohort study.

Numerous trials have shown that lowering blood pressure (BP) reduces cardiovascular risk and mortality, yet data about the impact of BP on cardiovascular death risk in patients aged ≥80 years with acute myocardial infarction (AMI) are sparse. This study explored the prognostic value of BP for cardiovascular death during the first 48 h after admission following AMI among patients aged ≥80 years. A total of 1005 patients ≥80 years with AMI were enrolled. Average BP parameters, including systolic, diastolic, and pulse BP, over the first 48 h after admission were calculated. The end point was cardiovascular death. Receiver operating curve (ROC) analysis was used to identify whether BP was relevant to cardiovascular death. The relationship between BP levels and cardiovascular death was evaluated by Cox regression models. ROC analysis showed that average diastolic blood pressure (aDBP), but not systolic and pulse BP, was relevant to cardiovascular death, and the optimal cutoff was 65 mmHg. During the 2.9-year follow-up, patients who died from a cardiovascular cause had lower aDBP levels than those who did not (p = 0.002). Patients with aDBP <65 mmHg had a 1.5-fold higher incidence of cardiovascular death than those with aDBP ≥65 mmHg (35.9% vs. 24.0%; p < 0.001). In multivariable regression analysis, low aDBP remained a strong and independent predictor of cardiovascular death (adjusted hazard ratio 1.907; 95% CI 1.303-2.792). aDBP was independently associated with cardiovascular death in patients aged ≥80 years with AMI, suggesting that aDBP may be a useful index to predict worse outcome in these patients.

Correlation Analysis of Huayu Tongmai Decoction Intervention and Prognosis Indexes of Patients with Carotid Atherosclerosis.

Background: Carotid atherosclerosis (CAS) is a common disease which seriously threatens the health of senile patients. The studies have indicated that traditional Chinese medicine (TCM) may effectively improve the symptom of CAS, while the therapeutic effect of Huayu Tongmai decoction on CAS remains unclear. Thus, this study aimed to explore the correlation between traditional Chinese medicine Huayu Tongmai decoction intervention and prognosis indexes of patients with CAS. Methods: Ninety CAS patients admitted to Zibo TCM-Integrated Hospital from September 2018 to September 2020 were selected as the research object and randomly divided into the control group and the observation group according to the male-female ratio of 1 : 1. Patients in the control group accepted the atorvastatin intervention, and on this basis, patients in the observation group were further intervened with TCM Huayu Tongmai decoction. Before and after treatment, patients' levels of total cholesterol (TC), triglyceride (TG), and low-density lipoprotein cholesterol (LDL-C) were measured by the enzyme photometric colorimetry; hypersensitive c-reactive protein (hs-CRP) levels were measured by the ELISA method; nitric oxide (NO) levels were measured by the nitrate reductase assay and endothelin-1 (ET-1) levels were measured by radioimmunoassay; and the right and left carotid internal diameter (CAD), intima-media thickness (IMT), and plaque volume were measured by carotid ultrasonography. Results: The TC, TG, and LDL-C levels significantly decreased in patients compared to those before intervention; compared with the control group, patients who accepted Huayu Tongmai decoction combined with atorvastatin saw more significant improvement in their blood lipid indexes (P < 0.01); after intervention, patients' hs-CRP and ET-1 levels dropped significantly while the NO level rose remarkably, and between the two groups, the improvement in levels of hs-CRP, ET-1, and NO of patients in the observation group was significantly better (P < 0.01); it was concluded from the imaging diagnosis results that compared with using atorvastatin alone, the combined intervention could better improve patients' CAD, IMT, and plaque volume. Conclusion: Huayu Tongmai decoction can effectively improve patients' blood lipid, reduce inflammatory response, enhance levels of relevant regulatory factors of CAS, and alleviate the symptoms.

Evaluation of the Antidepressant Effect of the Functional Beverage Containing Active Peptides, Menthol and Eleutheroside and Investigation of Its Mechanism of Action in Mice.

RESEARCH BACKGROUND: Depression has become a global threat to human health. In order to solve it, researchers have conducted multi-faceted studies including diet. Many food-derived bioactive substances have shown antidepressant effects. However, there are few studies on the design of industrialized food with antidepressant effect. This study aims to evaluate the antidepressant effect of a functional beverage made from several ingredients with potential antidepressant function and investigate its antidepressant mechanisms. EXPERIMENTAL APPROACH: The beverage consists of peppermint oil, active peptides derived from bovine milk casein and Acanthopanax senticosus extract (ASE) whose active ingredient is eleutheroside. Different amounts of ASE were evaluated to determine the optimal concentration of eleutheroside in this functional beverage to deliver the best antidepressant effect through extensive behavioral testing, including preliminary acute stress experiments and further chronic unpredictable mild stress test. RESULTS AND CONCLUSIONS: The results demonstrated that the beverage with 15 mg/kg of eleutheroside could significantly reduce the mice's immobility time of tail suspension test and forced swimming test, recover mice's sucrose preference and behavior changes in the open field test, improve the contents of dopamine, norepinephrine, 5-hydroxytryptamine and the activity of superoxide dismutase and reduce the content of malondialdehyde in mice's brains, which indicated that the improvement of monoamine neurotransmitter systems and antioxidation was one potential mechanism of antidepressant action. NOVELTY AND SCIENTIFIC CONTRIBUTION: This study provides a design of antidepressant functional beverage and an efficient way for the prevention and treatment of depression.

Differential leukocyte counts and cardiovascular mortality in very old patients with acute myocardial infarction: a Chinese cohort study.

BACKGROUND: Total leukocyte and differential Leukocyte counts are prognostic indictors in patients with coronary artery disease (CAD). However, there is no data available regarding their prognostic utility in very old patients with acute myocardial infarction (AMI). The aim of this study is to investigate the potential role of different leukocyte parameters in predicting the mortality among very old patients with AMI. METHODS: A total of 523 patients aged over 80 years with AMI were consecutively enrolled into this study. Leukocyte and its subtypes were obtained at admission in each patient. The primary study endpoint was cardiovascular mortality. Patients were followed up for an average of 2.2 years and 153 patients died. The associations of leukocyte parameters with mortality were assessed using Cox regression analyses. The concordance index was calculated to test the model efficiency. RESULTS: In multivariable regression analysis, neutrophils-plus-monocytes-to-lymphocytes ratio (NMLR) and neutrophils-to-lymphocytes ratio (NLR) were two most significant predictors of mortality among all the leukocyte parameters (HR = 3.21, 95% CI 1.75-5.35; HR = 2.79, 95% CI 1.59-4.88, respectively, all p < 0.001, adjusted for age, male gender, body mass index, family history of CAD, smoking, hypertension, diabetes mellitus, high-density lipoprotein cholesterol (HDL-C), non-HDL-C, high sensitivity C-reactive protein, creatinine, left ventricular ejection fraction, troponin I, use of statin, angiotensin-converting enzyme inhibitors/angiotensin receptor blockers, and percutaneous coronary intervention). Furthermore, adding NMLR and NLR into the Cox model increased the C-statistic by 0.038 and 0.037 respectively, which were more significant than that of other leukocyte parameters. Besides, addition of NMLR and NLR to the Canada Acute Coronary Syndrome Risk Score model also increased the C-statistic by 0.079 and 0.077 respectively. CONCLUSION: Our data firstly indicated that most leukocyte subtypes were independent markers for the mortality in very old patients with AMI, while NMLR and NLR appeared to be more effective.

Lipoprotein(a) and cardiovascular death in oldest-old (≥80 years) patients with acute myocardial infarction: A prospective cohort study.

BACKGROUND AND AIMS: Compared with what is known about the prognostic value of lipoprotein(a) [Lp(a)] in middle-aged patients with atherosclerotic cardiovascular disease (ASCVD), less is understood concerning the role of Lp(a) in oldest-old (≥80 years old) with ASCVD. The aim of the present study was to investigate the relationship between Lp(a) and cardiovascular death (CD) among the oldest-old with acute myocardial infarction (AMI). METHODS: A total of 1008 patients with AMI, older than 80 years, were consecutively enrolled between January 2012 and August 2018. The clinical characteristics were collected and Lp(a) concentrations were measured by the immunoturbidimetric method at baseline. The relationship between plasma Lp(a) concentration (≤10 mg/dL, 10-30 mg/dL, >30 mg/dL) and CD was evaluated by Kaplan-Meier analysis and Cox proportional hazard models. RESULTS: During an average of 36.26 months of follow-up, 287 CD occurred. Data showed that patients with high Lp(a) levels (>30 mg/dL) had the highest rate of CD (p < 0.05). Kaplan-Meier analysis showed that the high Lp(a) group had the lowest event-free survival rate in the oldest-old with AMI (p = 0.030). In addition, subjects with Lp(a) > 30 mg/dL had a 1.5-fold (95% confidence interval: 1.083-2.132) higher risk of CD compared with those with Lp(a) ≤10 mg/dL in fully adjusted Cox proportional hazards model. CONCLUSIONS: The current data firstly showed that plasma Lp(a) concentration was associated with the risk of CD in oldest-old with AMI, suggesting that Lp(a) could be a useful adjunctive measurement in the evaluation of CD in this population.

Lipoprotein(a) Is Associated with the Presence and Severity of New-Onset Coronary Artery Disease in Postmenopausal Women.

Background: Lipid disorder was one of the major risk factors for coronary artery disease (CAD), especially in postmenopausal women, whose lipid profile significantly changed during the transition period to menopause. The aim of the present study was to examine whether plasma lipoprotein(a) [Lp(a)] was a biomarker for predicting the presence and severity of CAD in postmenopausal women. Methods: A total of 783 postmenopausal women who had their first angina-like chest pain were enrolled and classified into two groups according to the results of coronary angiography: CAD group (n = 309) and age-matched non-CAD group (n = 309). Patients with CAD were further divided into the three groups based on Gensini score (GS). The relationships of plasma Lp(a) levels to the presence and severity of CAD were evaluated, and the predictive value of Lp(a) for CAD was also examined. Results: CAD group had higher Lp(a) levels when compared to non-CAD ones (p < 0.001). The multivariate logistic regression analysis suggested that Lp(a) was an independent predictor for the presence of CAD (p < 0.001). Plasma levels of Lp(a) were significantly related to GS (p < 0.001). In addition, plasma Lp(a) level was significantly elevated according to the tertiles of GS (p = 0.001) and was independently associated with high GS (p < 0.001). In receiver-operating characteristic analysis for predicting the presence of CAD in postmenopausal women, Lp(a) was found to have the area under the curve of 0.703, with an optimal cutoff value of 255.69 mg/L. Conclusions: Lp(a) is an independent risk factor for predicting the presence and the severity of new-onset CAD in postmenopausal women, suggesting that Lp(a) may be a lipid target for prevention and treatment in such patients.

Associations between Self-Determined Motivation, Accelerometer-Determined Physical Activity, and Quality of Life in Chinese College Students.

Purpose: To better promote college students' physical activity (PA) and quality of life (QoL), it is imperative to understand this population's PA correlates, such as self-determined motivation and perceived competence. However, few studies existed in this area of inquiry among Chinese college students. Thus, the purpose of this study was to examine the relationships among Chinese college students' self-determined motivation, PA, and QoL. Method: A total of 220 college students (115 females; Mage = 20.29 years, SD = 2.37; MBMI = 20.67) were recruited from one university in south-central China. Participants were instructed to wear the ActiGraph GT9X Link (ActiGraph, Pensacola, FL, USA) accelerometers for 7 days. A minute-by-minute stepping rate methodology was used to determine participants sedentary behaviors, light physical activity (LPA) and moderate-to-vigorous physical activity (MVPA). Participants' self-determined motivation (autonomous, controlled, and amotivation), perceived competence, and QoL (physical function, stress, depression, fatigue, sleep, and social issues) were assessed by a battery of validated surveys in June 2017. Results: Participants reported moderate-high levels of PA correlates and QoL as the means ranged from 5.5 to 6 (out of 7) for PA correlates and 2.75 to 4 (out of 4) for QoL. The minute-by-minute stepping rate revealed participants had average 580.51 min/day in sedentary, 134.77 min/day in LPA, and 1.57 min/day in MVPA. Regression analyses for physical function, stress, depression, and social issues suggested that the models explained 4%-8% of the variances. Specifically, perceived competence was the negative predictor of the problems with physical function (ß = -0.17, p < 0.05) and depression (ß = -0.18, p < 0.01), amotivation was positively associated with depression and stress (p < 0.05). Additionally, controlled motivation predicted the ability to participate in social roles and activities (ß = 0.22, p < 0.05). No significant predictors emerged for fatigue or for sleep. Conclusions: Findings suggest Chinese college students' perceived competence and social support are critical for improving PA and QoL. In addition, strategies are needed to motivate Chinese college students to engage in PA participation and improve overall well-being.

A nomogram for determining the disease-specific survival in Ewing sarcoma: a population study.

BACKGROUND: We aimed to develop and validate a nomogram for predicting the disease-specific survival of Ewing sarcoma (ES) patients. METHODS: The Surveillance, Epidemiology, and End Results (SEER) program database was used to identify ES from 1990 to 2015, in which the data was extracted from 18 registries in the US. Multivariate analysis performed using Cox proportional hazards regression models was performed on the training set to identify independent prognostic factors and construct a nomogram for the prediction of the 3-, 5-, and 10-year survival rates of patients with ES. The predictive values were compared by using concordance indexes (C-indexes), calibration plots, integrated discrimination improvement (IDI), net reclassification improvement (NRI), and decision curve analysis (DCA). RESULTS: A total of 2,643 patients were identified. After multivariate Cox regression, a nomogram was established based on a new model containing the predictive variables of age, race, extent of disease, tumor size, and therapy of surgery. The new model provided better C-indexes (0.684 and 0.704 in the training and validation cohorts, respectively) than the model without therapy of surgery (0.661 and 0.668 in the training and validation cohorts, respectively). The good discrimination and calibration of the nomogram were demonstrated for both the training and validation cohorts. NRI and IDI were also improved. Finally, DCA demonstrated that the nomogram was clinically useful. CONCLUSION: We developed a reliable nomogram for determining the prognosis and treatment outcomes of patients with ES in the US. However, the proposed nomogram still requires external data verification in future applications, especially for regions outside the US.

Development and validation of a nomogram for osteosarcoma-specific survival: A population-based study.

This study aimed to establish a comprehensive prognostic system for osteosarcoma based on a large population database with high quality.The Surveillance, Epidemiology, and End Results (SEER) Program database was used to identify patients with osteosarcoma from 1973 to 2015. Multivariate analysis was performed to screen statistically significant variables. A nomogram was constructed by R software to predict the 3-, 5- and 10-year survival rates. Predictive abilities were compared by C-indexes, calibration plots, integrated discrimination improvement (IDI), net reclassification improvement (NRI), as well as decision curve analysis (DCA).In total, 4505 osteosarcoma patients were identified. They were divided into training (70%, n = 3153) and validating (30%, n = 1352) groups. Multivariate analyses identified independent predictors. Subsequently, the nomogram system of a new model was established, which comprised 7 variables as age, sex, site, decade of diagnosis (DOD), extent of disease (EOD), tumor size and patients undergoing tri-modality therapy (surgery, radiotherapy and chemotherapy). It provided better C-indexes than the model without therapies (0.727, 0.712 vs 0.705, 0.668) in the 2 cohort, respectively. As well, the new model had good performances in the calibration plots. Moreover, both IDI and NRI improved for 3-, 5- and 10-year follow-up of C-indexes. Finally, DCA demonstrated that the nomogram of new model was clinically meaningful.We developed a reliable nomogram for prognostic determinants and treatment outcome analysis of osteosarcoma, thus helping better choose medical examinations and optimize therapeutic regimen under the cooperation among oncologists and surgeons.

Probiotic Bacillus amyloliquefaciens C-1 Improves Growth Performance, Stimulates GH/IGF-1, and Regulates the Gut Microbiota of Growth-Retarded Beef Calves.

Growth retardation of calves is defined as a symptom of impaired growth and development, probably due to growth hormone disorder as well as natural and environmental factors in livestock. The growth-promoting effects of probiotics were determined in 50 growth-retarded growth calves. They were supplied with Bacillus amyloliquefaciens C-1 (Ba, 4 × 1010CFU/d, n = 16), B. subtilis (Bs, 4 × 1010CFU/d, n = 18), and negative control (NC, n = 16) for 30 days. Pre- and post-intervention, the growth performance (weight gain rate, feed intake and feed conversion rate) was analyzed, the serum GH, IGH-1 and immunoglobulin levels were assayed, and the fecal microbiota was detected. Calves in Ba and Bs groups demonstrated increased body weight gain, feed intake and GH/IGF-1 levels, as well as a more efficient feed conversion rate, compared with NC group (P < 0.05). Additionally, the abundances of bacteria contributing to the production of energy and SCFAs (short chain fatty acids), including Proteobacteria, Rhodospirillaceae, Campylobacterales, and Butyricimonas were increased compared with NC group (P < 0.05, FDR < 0.1); and the suspected pathogens, which included Anaeroplasma and Acholeplasma were decreased (P < 0.05, FDR < 0.1) in both the Bs and Ba groups. Akkermansia, which is involved in the intestinal mucosal immune response, was increased in Bs group after intervention (P < 0.05, FDR < 0.1), but exhibited no obvious difference in Ba group. The increased bacterial genera in Ba group were Sphaerochaeta and Treponema (P < 0.05, FDR < 0.1). These results indicate that the probiotics B. amyloliquefaciens and B. subtilis exhibited similar therapeutic potential in terms of growth performance by regulating hormones, and improving the intestinal and rumen development in growth-retarded animals.

miR-644a Inhibits Cellular Proliferation and Invasion via Suppression of CtBP1 in Gastric Cancer Cells.

Epithelial-mesenchymal transition (EMT) is one of the most important mechanisms in the metastasis of various cancers, including gastric cancer (GC). In this study, we explored the putative significance of miR-644a and its role in EMT-mediated metastasis of GC. We first detected the expression of miR-644a in a cohort of 107 GC tissues using quantitative RT-PCR. The expression of miR-644a was suppressed in GC tissues and was associated with a later clinical stage and tumor metastasis. Restoring the expression of miR-644a could significantly suppress the migration and invasion of HGC-27 and SGC-7901 cells, which might be correlated to its suppressive effect on the EMT process. We also found that carboxyl-terminal-binding protein 1 (CtBP1) was a putative target gene of miR-644a in GC and might be involved in the suppressive effect. Collectively, through targeting CtBP1-mediated suppression of the EMT process, miR-644a might suppress the tumor metastasis of GC cells.

Sigma Factor Regulated Cellular Response in a Non-solvent Producing Clostridium beijerinckii Degenerated Strain: A Comparative Transcriptome Analysis.

Clostridium beijerinckii DG-8052, derived from NCIMB 8052, cannot produce solvent or form spores, a phenomenon known as degeneration. To explore the mechanisms of degeneration at the gene level, transcriptomic profiles of the wild-type 8052 and DG-8052 strains were compared. Expression of 5168 genes comprising 98.6% of the genome was assessed. Interestingly, 548 and 702 genes were significantly up-regulated in the acidogenesis and solventogenesis phases of DG-8052, respectively, and mainly responsible for the phosphotransferase system, sugar metabolic pathways, and chemotaxis; meanwhile, 699 and 797 genes were significantly down-regulated, respectively, and mainly responsible for sporulation, oxidoreduction, and solventogenesis. The functions of some altered genes, including 286 and 333 at the acidogenesis and solventogenesis phases, respectively, remain unknown. Dysregulation of the fermentation machinery was accompanied by lower transcription levels of glycolysis rate-limiting enzymes (pfk and pyk), and higher transcription of cell chemotaxis genes (cheA, cheB, cheR, cheW, and cheY), controlled mainly by σ54 at acidogenesis. Meanwhile, abnormal spore formation was associated with repressed spo0A, sigE, sigF, sigG, and sigK which are positively regulated by σ70, and correspondingly inhibited expression of CoA-transferase at the solventogenesis phase. These findings indicated that morphological and physiological changes in the degenerated Clostridium strain may be related to altered expression of sigma factors, providing valuable targets for strain development of Clostridium species.

Predictive value of non-fasting remnant cholesterol for short-term outcome of diabetics with new-onset stable coronary artery disease.

BACKGROUND: The relationship between non-fasting remnant cholesterol and cardiovascular outcome in the era of potent statin therapy remained to be elucidated. METHODS: A cohort study of three hundred and twenty eight diabetics diagnosed with new-onset stable coronary artery disease (CAD) by coronary angiography were enrolled. All cases were followed up for an average duration of twelve months. The association between baseline remnant cholesterol levels and major cardiovascular outcomes were evaluated using the receivers operating characteristic (ROC) curves and Cox proportional hazards regression analysis. RESULTS: During a period of 12-month's follow-up, 14.3% patients (47/328) underwent pre-specified adverse outcomes. The remnant cholesterol associated with high sensitivity C-reactive protein, neutrophil count and fibrinogen (R 2 = 0.20, 0.12 and 0.14; P = 0.000, 0.036 and 0.010 respectively). Area under the ROC curves (AUC) indicated discriminatory power of the remnant cholesterol to predict the adverse outcomes for this population (AUC = 0.64, P < 0.005). Kaplan-Meier curve suggested that the lower levels of remnant cholesterol showed relatively lower cardiac events for diabetic patients with stable CAD (Log rank X 2 = 8.94, P = 0.04). However, according to multivariate Cox proportional hazards regression, apart from hemoglobin A1C (Hazard ratio [H.R.] =1.38, 95% CI: 1.14-1.66, P = 0.001) and Gensini scores (H.R. = 1.00, 95% CI: 1.00-1.02; P = 0.035), remnant cholesterol did not qualify as an independent predictor of adverse prognosis in these settings (H.R. = 1.05, 95% CI: 0.46-2.37, P = 0.909). CONCLUSIONS: Non-fasting remnant cholesterol was associated with inflammatory biomarkers and high incidence of revascularization, but not qualified as an independent predictor for short-term prognosis of diabetics with new-onset stable coronary artery disease.

Is the ratio of apoB/apoA-1 the best predictor for the severity of coronary artery lesions in Chinese diabetics with stable angina pectoris? An assessment based on Gensini scores.

BACKGROUND: There is a paucity of data about the best lipid ratio predicting the severity of coronary artery disease (CAD) in patients with diabetes mellitus. We determined the relationship between five conventional lipid ratios and the extent of coronary artery lesions in Chinese Type 2 diabetics with stable angina pectoris (SAP). METHODS: A prospective cohort study within 373 type 2 diabetic patients diagnosed with stable CAD by coronary angiography was performed. All patients were classified into three groups according to the tertiles of Gensini scores (GS, low group < 8 points n = 143; intermediate group 8-28 points, n = 109; high group > 28 points, n = 121). Association between the ratios of apolipoprotein (apo) B and apoA-1, total cholesterol and high density lipoprotein cholesterol (TC/HDL-C), triglycerides and HDL-C (TG/HDL-C), low density lipoprotein cholesterol and HDL-C (LDL-C/HDL-C), Non-HDL-C/HDL-C and GS were evaluated using the receivers operating characteristic (ROC) curves and multivariate logistic regression models. RESULTS: The ratio of apoB/apoA-1, TC/HDL-C, LDL-C/HDL-C, and Non-HDL-C/HDL-C were correlated with Gensini scores. Area under the ROC curves for predicting high Gensini scores in the ratios of apoB/apoA-1, TC/HDL-C, LDL-C/HDL-C and Non-HDL-C/HDL-C were 0.62, 0.60, 0.59 and 0.60, respectively (P < 0.005 for all). According to multivariate logistic regression analysis after adjusted with demographic characteristic and other lipid parameters, the ratio of apoB/apoA-1 is qualified as an independent discriminator for the severity of CAD. However, after further adjusting different baseline variables, such as left ventricular ejective fraction, hemoglobin A1c, leukocytes count and serum creatinine, none of the above lipid ratios remained. CONCLUSIONS: Compared with other lipid parameters, the ratio of apoB/apoA-1 appears to be more significantly correlated with the extent of coronary artery lesions in Chinese diabetics, but it was not an independent predictor in these settings.

Impact of 1, 25-(OH)2D3 on Left Ventricular Hypertrophy in Type 2 Diabetic Rats.

OBJECTIVE: To investigate the impact of 1, 25-(OH)2D3 on left ventricular hypertrophy (LVH) in type 2 diabetic rats. METHODS: Type 2 diabetic mellitus (DM) model rats were established by intraperitoneally injecting with 30 mg/kg streptozotocin. After 8 weeks, 19 male rats were identified as diabetic with left ventricular hypertrophy (LVH) by ultrasound examination, and randomly assigned into three groups: untreated (DM-LVH, n=7), treated with insulin (DM-LVH+INS, n=6), and treated with 1, 25-(OH)2D3 (DM-LVH+VD, n=6). Healthy male rats were used as the controls group (n=6). The fasting blood glucose and the insulin level were determined weekly. The left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor level were determined by 4 weeks later. RESULTS: In the DM-LVH model group, the insulin level was significantly decreased compared with the non-diabetic control group (P<0.05), whereas the blood glucose, left ventricular mass index, myocardial collagen content, collagen volume fraction, and 1, 25-(OH)2D3-receptor expression were significantly increased (all P<0.05). In the DM-LVH+INS and DM-LVH+VD groups, the insulin levels were significantly increased compared with the DM-LVH model group (P<0.05), whereas the other parameters were significantly decreased (all P<0.05). CONCLUSION: 1, 25-(OH)2D3 could reverse LVH in diabetic rats and that the mechanism may involve stimulating insulin secretion and reducing blood glucose via direct up-regulation of 1, 25-(OH)2D3-receptor expression.

[Effects of telmisartan on endoplasmic reticulum stress induced cardiomyocyte apoptosis in abdominal aortic banded rats].

OBJECTIVE: To investigate the effects of telmisartan on endoplasm reticulum (ER) stress signal pathways and cardiomyocyte apoptosis in abdominal aortic banded rats. METHODS: Male SD rats were randomly divided into sham-operated group, abdominal aortic banding group (AAB) and AAB + telmisartan (5 mgxkg(-1)xd(-1) per gavage, beginning at 1 week before AAB for 8 weeks, n = 10 each). Ten weeks post AAB, hemodynamic measurements were performed, whole heart and left ventricular weights were obtained, cardiomyocyte apoptosis was measured by TUNEL method. Myocardial GRP78 and CHOP protein expressions were detected by Western blot and immunohistochemistry. RESULTS: The ratio of left ventricular weight to body weight, the ratio of heart weight to body weight, left ventricular end diastolic pressure and the apoptosis index were significantly increased while left ventricular end systolic pressure and +/- dp/dt(max) were significantly decreased in AAB group than those in sham-operated group (all P < 0.01), these changes could be significantly attenuated by telmisartan (all P < 0.01). Moreover, myocardial GRP78 and CHOP expressions were significantly upregulated in AAB group than those in sham-operated group and telmisartan could significantly downregulate the increased GRP78, CHOP expressions (all P < 0.01). CONCLUSIONS: Increased ER stress might be responsible for enhanced cardiomyocyte apoptosis in AAB rats. Telmisartan effectively attenuated the cardiomyocyte apoptosis and cardiac hypertrophy in AAB rats possibly through reducing ER stress.